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. 2020 Feb 4;48(6):3071–3088. doi: 10.1093/nar/gkaa055

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Negative growth regulation by a doxycycline-controlled glutamine tRNA synthetase gene GLN4. (A) The growth of a wild-type strain on a range of doxycycline-containing agar was compared to that of a tet-off GLN4 integrant, and a tet-off GLN4 integrant C-terminally tagged with HA epitope. (B) Using the engineered C-terminal epitope tag on Gln4p, Western blot band intensities were quantitatively compare to an HA-tagged Gln4p standard expressed in E. coli, using a standard curve (not shown). (C) The growth rate constant for a tet-off GLN4 integrant C-terminally tagged with HA epitope was measured across a range of doxycycline concentrations. (D) Using quantitative Western blots, the cellular content of Gln4p was calculated allowing growth rate to be fitted biphasically to Gln4 content. The vertical dashed line indicates the cellular content of Gln4p estimated in a proteomic meta-study (58).